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Determining Membrane Protein Topology Using Fluorescence Protease Protection (FPP)

17.6K Views

08:14 min

April 20th, 2015

April 20th, 2015

17,587 Views

0:05

Title

1:42

Transfection of Cells and Cell Culture

3:02

Establishing Optimal Conditions for Plasma Membrane Permeabilization

4:59

Protease Treatment of Fluorescent Proteins and Image Analysis

5:56

Results: Analysis of Protein Topology Using FPP

7:48

Conclusion

Here, we present a protocol to determine the orientation and topology of integral membrane proteins in living cells. This simple protocol relies on selective protease sensitivity of chimeras between the protein of interest and GFP.

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